The Core will provide facilities for the application of molecular biology, immunocytochemical and imaging techniques. To facilitate these goals the core will provide: 1) technical assistance for tissue preparation and cell culture for molecular biology techniques and immunohistochemistry, 2) provide technical assistance in specialized biochemical or immunohistochemical techniques, 3) ready access to specialized equipment, 4) cost effectiveness through centralized equipment and facilities, 5) sharing of knowledge between different investigators. The services, instruction and specialized equipment provided by the Core will play a critical role in the successful completion of the research projects in an efficient and effective manner. The specific aims of the Molecular Biology, Immunocytochemistry and Imaging Core are: 1. To provide assistance and facilities to investigators for cell culture, gene transfer in cultured cells and in tissues, and help in the quantification of gene expression by Northern and Western blot analysis and to estimate the functionality of the transferred gene using appropriate biochemical assays. 2. To provide facilities for histochemical and fluorescence applications for the detection of reporter proteins and transgene products in cells and tissues. 3. To provide assistance in immunofluorescence determinations of cell type and nerve terminals specific transgene expression using multiple antibodies. 4. To actively collaborate with the PPG investigators in designing and executing experiments pertaining to measurements of intracellular cation homeostasis, membrane potential and redox state of the cell during stimulus-response coupling. This will generally involve procedures of microscopic video digital image analysis system, and confocal microscopy. 5. To provide facilities and assistance in molecular biology techniques for the estimation of mRNA levels by Northern analysis and RT-PCR. 6. The activities of the core will be directed towards fostered development of new technology as well as collaborative investigations of focused problems.